DLGH1 AND UROGENITAL DEVELOPMENT

DLGH1 和泌尿生殖系统发育

• 批准号: 8288317
• 负责人: JEFFREY H MINER
• 金额: $ 31.66万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-14 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8288317
• 关键词: Affect; Alleles; Animals; Binding Proteins; Biological Process; Biology; Cell Line; Cell Polarity; Cell Proliferation; Cells; Complex; Cytoskeleton; Data; Defect; Development; Drosophila And protein; Drosophila genus; Embryo; Employee Strikes; Epithelial; Epithelial Cells; Epithelium; Event; Exhibits; Eyelid structure; Family; Family member; Female; Fibroblasts; Gene Expression; Genes; Genetic; Genitourinary system; Golgi Apparatus; Guanylate kinase; Health; Homologous Gene; Human; Hydronephrosis; Immigration; Immunofluorescence Immunologic; In Vitro; Kidney; Kidney Diseases; Knock-out; Knockout Mice; Life; MAPK14 gene; Mammalian Cell; Mediating; Mesenchymal; Mesenchyme; Metanephric Diverticulum; Methods; Microtubule-Organizing Center; Molecular; Molecular Profiling; Morphogenesis; Morphology; Motion; Mus; Muscle; Mutagenesis; Mutant Strains Mice; Mutate; Mutation; Nature; Palate; Pattern; Peristalsis; Phenotype; Phosphotransferases; Play; Population; Prevention; Protein Binding Domain; Protein Family; Proteins; Regulation; Reporter; Research; Role; SH3 Domains; Scaffolding Protein; Seminal Vesicles; Signal Transduction; Site; Smooth Muscle; Smooth Muscle Myocytes; Staging; Sternum; Stromal Cells; Structure; Structure of mesonephric duct; Suppressor-Effector T-Lymphocytes; Synapses; System; Testing; Time; Tissues; Transfection; Transgenes; Transgenic Mice; Tumor Suppressor Proteins; Ureter; Urinary tract; Urine; Urothelial Cell; Urothelium; Vagina; Work; Wound Healing; base; cell motility; cell type; fly; glycogen synthase kinase 3 beta; human DLG1 protein; imaginal disc; in vivo; male; member; membrane-associated guanylate kinase; migration; mouse model; mutant; novel; null mutation; postsynaptic density protein; reproductive; research study; synaptic function; urinary tract obstruction

DESCRIPTION (provided by applicant): The long term objective of this research involves determining the mechanism whereby the mouse discs- large homolog 1 gene, Dlgh1, regulates urogenital development. DLGH1 is a scaffolding protein with multiple protein-protein interaction domains, including three PDZ domains and one SH3 domain. Dlgh1 is a homolog of the Drosophila discs-large gene, dlg, which is a founding member of the eponymous PDZ family of proteins and also of the MAGUK (membrane-associated guanylate kinase) family. Studies in Drosophila suggest that dlg is a tumor suppressor, because mutation of dlg causes overgrowth of imaginal discs. Studies in both fly and mammalian cells suggest that dlg and its homologs are also involved in establishing and/or maintaining epithelial cell polarity. We have studied mice lacking DLGH1 and found several urogenital defects, including sporadic renal agenesis, congenital hydronephrosis, and reproductive tract abnormalities. We have shown that hydronephrosis is associated with dramatic smooth muscle alignment defects; the normally circular muscle aberrantly aligns in the longitudinal direction. This greatly impairs the squeezing motion of peristalsis, such that urine is retained in the kidney. In addition, ureteric stromal cells, which normally lie between the urothelium and the smooth muscle layers and express Raldh2, are absent from Dlgh1 mutant ureters. The absence of these cells could also affect ureteric function, either directly or indirectly. Immunofluorescence studies show that DLGH1 is expressed strongly in the urothelium and weakly in other cells of the ureter. We will determine which cellular compartment must express Dlgh1 for normal smooth muscle cell alignment and for differentiation and/or migration of ureteric stromal cells using a new floxed Dlgh1 allele and appropriate Cre transgenic mice. In addition, we will generate chimeric mice to investigate whether DLGH1 acts in a non-cell autonomous fashion. Morphological and functional assessments will be performed to determine the cellular requirements for Dlgh1 expression. Next we will use genetic methods to investigate the hypothesis that the interaction of other DLGH family members with known or suspected DLGH1 binding proteins are compensating in part for the absence of DLGH1. Finally, we will use in vitro methods to define the role of DLGH1 in cell migration, polarization, and signaling. PUBLIC HEALTH RELEVANCE: Congenital urinary tract abnormalities are a relatively common health problem in the human population and are responsible for a significant number of cases of progressive renal disease. We have generated a novel mouse model of hydronephrosis (Dlgh1-/- mice) involving the misalignment of ureteric smooth muscle, which is commonly found in human cases of urinary tract obstruction. A better understanding of the biology of DLGH1 could have important implications for understanding and perhaps for treatment or prevention of urinary tract abnormalities in humans.

描述（由申请人提供）：本研究的长期目标涉及确定小鼠椎间盘大同源物 1 基因 Dlgh1 调节泌尿生殖发育的机制。 DLGH1 是一种具有多个蛋白质-蛋白质相互作用结构域的支架蛋白，包括三个 PDZ 结构域和一个 SH3 结构域。 Dlgh1 是果蝇圆盘大基因 dlg 的同源物，dlg 是同名 PDZ 蛋白家族和 MAGUK（膜相关鸟苷酸激酶）家族的创始成员。果蝇研究表明 dlg 是一种肿瘤抑制因子，因为 dlg 突变会导致成虫盘过度生长。对果蝇和哺乳动物细胞的研究表明，dlg 及其同系物也参与建立和/或维持上皮​​细胞极性。我们研究了缺乏 DLGH1 的小鼠，发现了多种泌尿生殖缺陷，包括偶发性肾发育不全、先天性肾积水和生殖道异常。我们已经证明，肾积水与严重的平滑肌排列缺陷有关。正常的圆形肌肉在纵向上异常排列。这极大地损害了蠕动的挤压运动，使得尿液滞留在肾脏中。此外，通常位于尿路上皮和平滑肌层之间并表达 Raldh2 的输尿管基质细胞在 Dlgh1 突变型输尿管中不存在。这些细胞的缺失也可能直接或间接影响输尿管功能。免疫荧光研究表明，DLGH1 在尿路上皮中强烈表达，而在输尿管其他细胞中表达较弱。我们将使用新的 floxed Dlgh1 等位基因和适当的 Cre 转基因小鼠来确定哪个细胞区室必须表达 Dlgh1，以实现正常平滑肌细胞排列以及输尿管基质细胞的分化和/或迁移。此外，我们将生成嵌合小鼠来研究 DLGH1 是否以非细胞自主方式发挥作用。将进行形态学和功能评估以确定 Dlgh1 表达的细胞需求。接下来，我们将使用遗传方法来研究以下假设：其他 DLGH 家族成员与已知或可疑的 DLGH1 结合蛋白的相互作用部分补偿了 DLGH1 的缺失。最后，我们将使用体外方法来定义 DLGH1 在细胞迁移、极化和信号传导中的作用。公共卫生相关性：先天性尿路异常是人群中相对常见的健康问题，是导致大量进行性肾病病例的原因。我们建立了一种新的肾积水小鼠模型（Dlgh1-/- 小鼠），该模型涉及输尿管平滑肌错位，这在人类尿路梗阻病例中很常见。更好地了解 DLGH1 的生物学特性可能对理解人类尿路异常以及治疗或预防人类尿路异常具有重要意义。

项目成果

Role of the polarity protein Scribble for podocyte differentiation and maintenance.

极性蛋白 Scribble 在足细胞分化和维持中的作用。

• 发表时间: 2012
• 影响因子: 3.7
• 作者: Hartleben, Björn;Widmeier, Eugen;Wanner, Nicola;Schmidts, Miriam;Kim, Sung Tae;Schneider, Lisa;Mayer, Britta;Kerjaschki, Dontscho;Miner, Jeffrey H;Walz, Gerd;Huber, Tobias B
• 通讯作者: Huber, Tobias B

The scaffolding protein synapse-associated protein 97 is required for enhanced signaling through isotype-switched IgG memory B cell receptors.

支架蛋白突触相关蛋白 97 是通过同种型转换 IgG 记忆 B 细胞受体增强信号传导所必需的。

• 发表时间: 2012
• 影响因子: 7.3
• 作者: Liu, Wanli;Chen, Elizabeth;Zhao, Xing Wang;Wan, Zheng Peng;Gao, Yi Ren;Davey, Angel;Huang, Eric;Zhang, Lijia;Crocetti, Jillian;Sandoval, Gabriel;Joyce, M Gordon;Miceli, Carrie;Lukszo, Jan;Aravind, L;Swat, Wojciech;Brzostowski, Joseph;Pierc
• 通讯作者: Pierc